Quality checks on the Salk polio vaccine at Glaxo's virus research laboratory in Buckinghamshire, UK, in January 1956.

How Are Vaccines Made?

BY Tara Smith

February 14, 2018

Quality checks on the Salk polio vaccine at Glaxo's virus research laboratory in Buckinghamshire, UK, in January 1956.

Photo by Fox Photos/Getty Images

Vaccines have long been hailed as one of our greatest public health achievements. They can be made to protect us from infections with either viral or bacterial microbes. Measles and smallpox, for example, are viruses; Streptococcus pneumoniae is a bacterium that causes a range of diseases, including pneumonia, ear and sinus infections, and meningitis. Hundreds of millions of illnesses and deaths have been prevented due to vaccines that eradicated smallpox and significantly reduced polio and measles infections. However, some misunderstanding remains regarding how vaccines are made, and why some scary-sounding ingredients [PDF] are included in the manufacturing process.

The production of our vaccines has greatly evolved since the early days, when vaccination was potentially dangerous. Inoculating an individual with ground-up smallpox scabs usually led to a mild infection (called "variolation"), and protected them from acquiring the disease the "regular" way (via the air). But there was always a chance the infection could still be severe. When Edward Jenner introduced the first true vaccination with cowpox, protection from smallpox became safer, but there were still issues: The cowpox material could be contaminated with other germs, and sometimes was transmitted from one vaccinated person to another, leading to the inadvertent spread of blood-borne pathogens. We’ve come far in the last 200 years.

There are different kinds of vaccines, and each requires different processes to move from the laboratory to your physician's office. The key to all of them is production of one or more antigens—the portion of the microbe that triggers a host immune response.

LIVE ATTENUATED VACCINES AND DEAD, "INACTIVATED" VACCINES

There are several methods to produce antigens. One common technique is to grow a virus in what's called a cell culture. Typically grown in large vats called bioreactors, living cells are inoculated with a virus and placed in a liquid growth medium that contains nutrients—proteins, amino acids, carbohydrates, essential minerals—that help the virus grow in the cells, producing thousands of copies of itself in each infected cell. At this stage the virus is also getting its own dose of protective medicine: antibiotics like neomycin or polymyxin B, which prevent bacterial and fungal contamination that could kill the cells serving as hosts for the virus.

Once a virus completes its life cycle in the host cell, the viruses are purified by separating them from the host cells and growth media, which are discarded. This is often done using several different types of filters; the viruses are small and can pass through holes in the filter that trap larger host cells and cell debris.

This is how "live attenuated vaccines" are created. These vaccines contain viruses that have been modified so that they are no longer harmful to humans. Some of them are grown for many generations in cells that aren't human, such as chicken cells, so that they have mutated to no longer cause harm to humans. Others, like the influenza nasal mist, were grown at low temperatures until they lost the ability to replicate in the warmer temperatures of the lungs. Many of these vaccines you were probably given as a child: measles, mumps, rubella ("German measles"), and chickenpox.

Live attenuated vaccines replicate briefly in the body, triggering a strong—and long-lasting—response from your immune system. Because your immune system kicks into high gear at what it perceives to be a major threat, you need fewer doses of the vaccine for protection against these diseases. And unlike the harmful form of the virus, it is extremely unlikely (because they only replicate at low levels) that these vaccines will cause the host to develop the actual disease, or to spread it to other contacts. One exception is the live polio vaccine, which could spread to others and, extremely rarely, caused polio disease (approximately one case of polio from 3 million doses of the virus). For this reason, the live polio virus was discontinued in the United States in 2000.

Scientists use the same growth technique for what are known as "killed" or "inactivated" vaccines, but they add an extra step: viral death. Inactivated viruses are killed, typically via heat treatment or use of a chemical such as formaldehyde, which modifies the virus's proteins and nucleic acids and renders the virus unable to replicate. Inactivated vaccines include Hepatitis A, the injected polio virus, and the flu shot.

A dead virus can't replicate in your body, obviously. This means that the immune response to inactivated vaccines isn't as robust as it is with live attenuated vaccines; replication by the live viruses alerts many different types of your immune cells of a potential invader, while killed vaccines primarily alert only one part of your immune system (your B cells, which produce antibodies). That's why you need more doses to achieve and maintain immunity.

While live attenuated vaccines were the primary way to make vaccines until the 1960s, concerns about potential safety issues, and the difficulty of making them, mean that few are attempting to develop new live attenuated vaccines today.

COMBINATION, BACTERIAL, AND GENETICALLY ENGINEERED VACCINES

Other vaccines aren't made of whole organisms at all, but rather bits and pieces of a microbe. The combination vaccine that protects against diphtheria, pertussis, and tetanus—all at once—is one example. This vaccine is called the DTaP for children, and Tdap for adults. It contains toxins (the proteins that cause disease) from diphtheria, pertussis, and tetanus bacteria that have been inactivated by chemicals. (The toxins are called "toxoids" once inactivated.) This protects the host—a.k.a. you, potentially—from developing clinical diphtheria and tetanus disease, even if you are exposed to the microorganisms. (Some viruses have toxins—Ebola appears to, for example—but they're not the key antigens, so they're not used for our current vaccines.)

As they do when developing live attenuated or inactivated vaccines, scientists who create these bacterial vaccines need some target bacteria to culture. But because the bacteria don't need a host cell to grow, they can be produced in simple nutrient broths by vaccine manufacturers. The toxins are then separated from the rest of the bacteria and growth media and inactivated for use as vaccines.

Similarly, some vaccines contain just a few antigens from a bacterial species. Vaccines for Streptococcus pneumoniae, Haemophilus influenzae type B, and Neisseria meningitidis all use sugars that are found on the outer part of the bacteria as antigens. These sugars are purified from the bacteria and then bound to another protein to enhance the immune response. The protein helps to recruit T cells in addition to B cells and create a more robust reaction.

Finally, we can also use genetic engineering to produce vaccines. We do this for Hepatitis B, a virus that can cause severe liver disease and liver cancer. The vaccine for it consists of a single antigen: the hepatitis B surface antigen, which is a protein on the outside of the virus. The gene that makes this antigen is inserted into yeast cells; these cells can then be grown in a medium similar to bacteria and without the need for cell culture. The hepatitis B surface antigen is then separated from the yeast and serves as the primary vaccine component.

OTHER INGREDIENTS IN VACCINES (AND WHY THEY'RE THERE)

Once you have the live or killed viruses, or purified antigens, sometimes chemicals need to be added to protect the vaccine or to make it work better. Adjuvants, such as aluminum salts, are a common additive; they help enhance the immune response to some antigens by keeping the antigen in contact with the cells of the immune system for a longer period of time. Vaccines for DTaP/Tdap, meningitis, pneumococcus, and hepatitis B all use aluminum salts as an adjuvant.

Other chemicals may be added as stabilizers, to help keep the vaccine working effectively even in extreme conditions (such as hot temperatures). Stabilizers can include sugars or monosodium glutamate (MSG). Preservatives can be added to prevent microbial growth in the finished product.

For many years, the most common preservative was a compound called thimerosal, which is 50 percent ethylmercury by weight. Ethylmercury doesn't stick around; your body quickly eliminates it via the gut and feces. (This is different from methylmercury, which accumulates in fish and can, at high doses, cause long-lasting damage in humans.) In 2001, thimerosal was removed from the vaccines given in childhood due to consumer concerns, but many studies have demonstrated its safety.

Finally, the vaccine is divided into vials for shipping to physicians, hospitals, public health departments, and some pharmacies. These can be single-dose or multi-dose vials, which can be used for multiple patients as long as they're prepared and stored away from patient treatment areas. Preservatives are important for multi-dose vials: bacteria and fungi are very opportunistic, and multiple uses increase the potential for contamination of the vaccine. This is why thimerosal is still used in some multi-dose influenza vaccines.

Though some of the vaccine ingredients sound worrisome, most of these chemicals are removed during multiple purification steps, and those that remain (such as adjuvants) are necessary for the vaccine's effectiveness, are present in very low levels, and have an excellent track record of safety.

Quality checks on the Salk polio vaccine at Glaxo's virus research laboratory in Buckinghamshire, UK, in January 1956.

The only thing you can be certain about in life is death. Or is it? Merriam-Webster defines death as "a permanent cessation of all vital functions." The Oxford English dictionary refines that to "the permanent ending of vital processes in a cell or tissue." But determining when someone is dead is surprisingly complicated—the medical definition has changed over the centuries and, in many ways, is still evolving.

DEATH, DEFINED

For most of human history, doctors relied on basic observations to determine whether or not a person had died. (This may be why so many feared being buried alive and went to great lengths to ensure they wouldn't be.) According to Marion Leary, the director of innovation research for the Center for Resuscitation Science at the University of Pennsylvania, "If a person wasn't visibly breathing, if they were cold and bluish in color, for example, they would be considered dead."

As time went on, the markers for death changed. Before the mid-1700s, for example, people were declared dead when their hearts stopped beating—a conclusion drawn from watching traumatic deaths such as decapitations, where the heart seemed to be the last organ to give up. But as our understanding of the human body grew, other organs, like the lungs and brain, were considered metrics of life—or death.

Today, that remains true to some degree; you can still be declared dead when your heart and lungs cease activity. And yet you can also be declared dead if both organs are still working, but your brain is not.

In most countries, being brain dead—meaning the whole brain has stopped working and cannot return to functionality—is the standard for calling death, says neuroscientist James Bernat, of the Geisel School of Medicine at Dartmouth College in New Hampshire. "A doctor has to show that the loss of brain function is irreversible," he tells Mental Floss. In some cases, a person can appear to be brain dead if they have overdosed on certain drugs or have suffered from hypothermia, for example, but the lack of activity is only temporary—these people aren't truly brain dead.

In the U.S., all states follow some form of the Uniform Determination of Death Act, which in 1981 defined a dead person as "an individual who has sustained either (1) irreversible cessation of circulatory and respiratory functions, or (2) irreversible cessation of all functions of the entire brain, including the brain stem."

But that's not the end of the story. In two states, New York and New Jersey, families can reject the concept of brain death if it goes against their religious beliefs. This makes it possible for someone to be considered alive in some states and dead in others.

A BLURRED LINE

In the past, if one of a person's three vital systems—circulation, respiration, and brain function—failed, the rest would usually stop within minutes of each other, and there was no coming back from that. But today, thanks to technological advances and medical breakthroughs, that's no longer necessarily the case. CPR can be performed to restart a heartbeat; a person who has suffered cardiac arrest can often be resuscitated within a 20- to 30-minute window (in rare cases, people have been revived after several hours). And since the 1950s, machines have been used to take on the role of many of the body's vital functions. People who stop breathing naturally can be hooked up to ventilators to move air in and out of their lungs, for example.

While remarkable, this life-extending technology has blurred the line between life and death. "A person can now have certain characteristics of being alive and others of being dead," Bernat says.

People with severe, irreversible brain damage fall into this mixed category. Many lie in intensive care units where ventilators breathe for them, but because they have minimal reflexes or movements, they're considered alive, especially by their families. Medical professionals, however, may disagree, leading to painful and complex debates about whether someone is alive.

Take the case of Jahi McMath, whose tonsil surgery in 2013, at age 13, went terribly wrong, leaving her brain dead—or so doctors thought. Her family refused to believe she was dead and moved her from Oakland, California, to New Jersey, where she was provided with feeding tubes in addition to her ventilator. After several months, her mother began recording videos that she said were proof that Jahi could move different parts of her body when asked to. Additional brain scans revealed that although some parts of her brain, like her brain stem, were largely destroyed, the structure of large parts of her cerebrum, which is responsible for consciousness, language, and voluntary movements, was intact. Her heart rate also changed when her mother spoke, leading a neurologist to declare last year, after viewing many of her mother's videos, that she is technically alive—nearly four years after she was pronounced brain dead. By her mother's reckoning, Jahi turned 17 on October 24, 2017.

Organ donation adds another layer of complications. Since an organ needs to be transplanted as quickly as possible to avoid damage, doctors want to declare death as soon as they can after a person has been disconnected from a machine. The protocol is usually to wait for five minutes after a donor's heart and breathing have stopped. However, some believe that's not long enough, since the person could still be resuscitated at that point.

Bernat—whose research interests include brain death and the definition of death, consciousness disorders including coma and vegetative states, and ethical and philosophical issues in neurology—disagrees. "I would argue that breathing and circulation has permanently ceased even if it hasn't irreversibly ceased," he says. "It won't restart by itself."

THE FUTURE OF BRINGING PEOPLE BACK TO LIFE

As resuscitation technology improves, scientists may find new ways to reverse death. One promising approach is therapeutic hypothermia. Sometimes used on heart attack patients who have been revived, the therapy uses cooling devices to lower body temperature, usually for about 24 hours. "It improves a patient's chance of recovering from cardiac arrest and the brain injury [from a lack of oxygen] that can result from it," says Leary, who specializes in research and education relating to cardiac arrest, CPR quality, and therapeutic hypothermia.

One more out-there possibility—which had its heyday in the early 2000s but still has its proponents today—is cryonic freezing, in which dead bodies (and in some cases, just people's heads) are preserved in the hope that they can be brought back once technology advances. Just minutes after death, a cryonaut's body is chilled; a chest compression device called a thumper keeps blood flowing through the body, which is then shot up with anticoagulants to prevent blood clots from forming; and finally, the blood is flushed out and replaced with a kind of antifreeze to halt the cell damage that usually occurs from freezing.

The idea is highly controversial. "It makes a good story for a movie, but it seems crazy to me," Bernat says. "I don't think it's the answer." But even if cryogenics is out, Bernat does believe that certain types of brain damage now thought to be permanent could one day be subject to medical intervention. "There is currently a huge effort in many medical centers to study brain resuscitation," he says.

Genetics provides another potential frontier. Scientists recently found that some genes in mice and fish live on after they die. And even more surprisingly, other genes regulating embryonic development, which switch off when an animal is born, turn on again after death. We don't yet know if the same thing happens in humans.

Quality checks on the Salk polio vaccine at Glaxo's virus research laboratory in Buckinghamshire, UK, in January 1956.

The word calorie carries a lot of weight. We know we're supposed to avoid too many of them, but things get more complicated after that. What, exactly, are calories, and how do I burn them?

THE SCIENCE OF THE CALORIE

A calorie is a unit of heat energy that fuels your body, making it possible to move, breathe, think, sleep—and even digest food to make more energy.

While there is some disagreement about who first coined the term calorie, we know the French chemist Antoine Lavoisier used it in experiments he conducted during the winter of 1782–1783. He used a device called a calorimeter to measure how much ice melted in a metal container due to the heat emitted by guinea pigs housed inside it. Over time, that measurement was refined by other scientists to mean the amount of energy needed to raise the temperature of a kilogram of water by 1°C—what's known as a kilocalorie.

The food calorie and a kilocalorie (kcal) are technically the same thing, but we use the term calorie rather than kilocalorie because of an American chemist named Wilbur Olin Atwater. In the late 1880s, Atwater traveled to Germany to study at physiologist Carl Voit's laboratory, where Voit was researching the nutritional value of food and animal feed. Inspired by that research, Atwater took measurements of different foods with a bomb calorimeter—a device that essentially measures the heat in food when burned—by having study participants eat, and then measuring and subtracting [PDF] the amount of heat leaving their bodies through respiration and waste. He used a respiration calorimeter to measure their breath and a bomb calorimeter to burn their poop, and from that calculated just how many calories were left in their bodies to be used. When writing about his research, Atwater used the word calorie (kcal wouldn't be used in America until 1894, when it was published in a physiology textbook).

Based on his experiments, Atwater created a system for calculating the calories that human bodies can get from our food. There are three types of food nutrients that deliver caloric energy—fats, proteins, and carbohydrates—and Atwater arrived at a caloric measurement of each: A fat gram has nine calories, while a gram of protein and a gram of carbohydrates each have four. That system was modified [PDF] by USDA scientists in 1973, but it's otherwise still the basis for how calories are calculated today.

WHAT HAPPENS TO A CALORIE IN YOUR BODY

When you eat, enzymes in the mouth, stomach, and intestine break down nutrients by turning fats into fatty acids, sugars into simple sugars, and proteins into amino acids. Then, using oxygen cells throughout your body, these components are broken down into energy—a process known as metabolism.

Most of the calories we burn each and every day are used just to keep our body functioning, with about half going toward powering our major organs—the brain, liver, kidneys, and heart. We use the rest for physical activity and the process of converting food to energy. Anything not used by the body is then stored, first in the liver and eventually as fat cells.

Some foods, like honey (carbohydrates), are easily digestible, whereas nuts (a mix of carbohydrates, fat, and protein) can't actually be fully digested at all. There are also digestibility differences within the same type of food. For example, in plants, older leaves tend to be sturdier (and therefore harder to digest) and less caloric than younger ones. Most significantly, especially in terms of human evolution, whenever we cook or process food, the body can get more calories as compared to that same food eaten raw. All of this has an impact on the amount of calories we can actually use.

There's no food you can eat to speed up the rate at which you burn calories (changes from foods like spicy peppers are fleeting), but factors like age and rapid, drastic weight loss can slow it down.

Building more muscle can increase your metabolic rate (although how much is debatable), since muscle requires more energy to function than fat does. And while cardiovascular exercise might not permanently boost your metabolism, it does burn calories; just how much depends on your weight and how vigorously you exercise.

Examples of higher calorie burning exercises include cycling and running, but almost every activity burns something, so you could potentially burn more calories throughout the day by consistently doing low-energy activities like gardening or pacing during a conference call than you would during 30 minutes of fast cycling.

CALORIES: A SCIENCE IN FLUX

We still use the Atwater system for calculating food calories, but it's far from perfect. For one thing, a USDA study found that people absorbed fewer calories from nuts than had been estimated under Atwater's system—a serving of almonds, for example, provided not 170 calories, but 129. There's some evidence that people tend to digest food at all sorts of different rates too, depending on the individual makeup of our gut bacteria, meaning that the absorption of calories may differ from person to person.

Scientists now believe the numbers on food labels are more of an estimate than a precise measurement. While companies are required to provide caloric information on food labels, the FDA doesn't specify exactly how those calories should be calculated. Some companies, like McDonald's, send their food to a lab for measurement, while others estimate the total by adding up the calorie count for each food component from the USDA's massive food composition database. As scientists continue to refine how we calculate calories, we'll come to have a better idea of the energy we can actually get from these different foods.